# Problems after trim.seqs

**URL:** https://forum.mothur.org/t/problems-after-trim-seqs/2081
**Category:** Commands in mothur
**Created:** [October 16, 2014, 7:42pm UTC](https://forum.mothur.org/t/problems-after-trim-seqs/2081 "2014-10-16T19:42:32Z")
**Posts on this page:** 7
**Page:** 1

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### Author: ![Lilianada](https://avatars.discourse-cdn.com/v4/letter/l/9de053/32.png) [@Lilianada](https://forum.mothur.org/u/Lilianada)
#### Post date: [October 16, 2014, 7:42pm UTC](https://forum.mothur.org/t/problems-after-trim-seqs/2081/1 "2014-10-16T19:42:32Z")

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Hi,  
I am very new to these analyses, I am following the 454 SOP protocol and I get an error message after this command:  
_\> summary.seqs(fasta=filename.shhh.trim.fasta, name=filename.shhh.trim.names)_  
which followed the trim.seqs command

_“trim.seqs(fasta=filename.shhh.fasta, name=filename.shhh.names, oligos=filename.oligos, pdiffs=2, bdiffs=1, maxhomop=8, minlength=200, flip=T, processors=2)”_

The error message says:  
[ERROR]: filename.shhh.trim.fasta is blank, aborting.  
Using filename.fasta as input file for the fasta parameter.  
[ERROR]: filename.shhh.trim.names is blank, aborting.

Using 2 processors.  
[WARNING]: This command can take a namefile and you did not provide one. The current namefile is filename.shhh.trim.names which seems to match filename.fasta.

When I checked the files created after the trim.seqs command I noticed that the _filename.shhh.trim.fasta_ and _filename.shhh.trim.names_ files are zero bytes

Could you help me identify the problem?

Thank you

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### Author: ![westcott](https://yyz2.discourse-cdn.com/flex036/user_avatar/forum.mothur.org/westcott/32/18_2.png) [@westcott](https://forum.mothur.org/u/westcott)
#### Post date: [October 16, 2014, 8:56pm UTC](https://forum.mothur.org/t/problems-after-trim-seqs/2081/2 "2014-10-16T20:56:14Z")

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This actually looks like an issue with trim.flows scrapping all your sequences. Here’s a link you may find helpful, [http://ωww.mothur.org/forum/viewtopic.php?f=3&t=2681](http://xn--ww-dcc.mothur.org/forum/viewtopic.php?f=3&t=2681).

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### Author: ![Lilianada](https://avatars.discourse-cdn.com/v4/letter/l/9de053/32.png) [@Lilianada](https://forum.mothur.org/u/Lilianada)
#### Post date: [October 17, 2014, 10:02pm UTC](https://forum.mothur.org/t/problems-after-trim-seqs/2081/3 "2014-10-17T22:02:45Z")

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Thank you,  
That helped me, I found out that it was the order=B that resulted in those zero byte files.

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### Author: ![wielern](https://avatars.discourse-cdn.com/v4/letter/w/cdc98d/32.png) [@wielern](https://forum.mothur.org/u/wielern)
#### Post date: [November 5, 2014, 4:16pm UTC](https://forum.mothur.org/t/problems-after-trim-seqs/2081/4 "2014-11-05T16:16:44Z")

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Hi,  
I ran trim.seqs and my trim file result was 0 bytes. i don’t have my flow data so I can’t run trim.flow. Is the problem is with my oligos file?

thank you,  
nimrod

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### Author: ![westcott](https://yyz2.discourse-cdn.com/flex036/user_avatar/forum.mothur.org/westcott/32/18_2.png) [@westcott](https://forum.mothur.org/u/westcott)
#### Post date: [November 5, 2014, 10:04pm UTC](https://forum.mothur.org/t/problems-after-trim-seqs/2081/5 "2014-11-05T22:04:02Z")

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It could be. Have you looked at the scrap codes mothur is giving you?

k linker missing  
b barcode missing  
s spacer missing  
f forward primer missing  
r reverse primer missing  
t too many differences to the linker, spacer, barcode, and forward primer  
l too short or too long  
h homopolymer length too long  
n too many ambiguous bases

Also, if a barcode is missing, then you’ll also be missing the forward primer and the spacer if you’ve included one.

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### Author: ![wielern](https://avatars.discourse-cdn.com/v4/letter/w/cdc98d/32.png) [@wielern](https://forum.mothur.org/u/wielern)
#### Post date: [November 9, 2014, 3:01pm UTC](https://forum.mothur.org/t/problems-after-trim-seqs/2081/6 "2014-11-09T15:01:50Z")

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Hi,

thank you, according to the scrap file I am missing the barccode and forward primer. how can I fix it (I attached below my oligos file)?

forward AGAGTTTGATCMTGGCTCAG 27f  
barcode TTGTTGCTGT Osnat1  
barcode GTGTGGTTGT Osnat2  
barcode TAGGTGGAAT Osnat3  
barcode TGTAGGTGGA Osnat4  
barcode TTAGTGGTGA Osnat5  
barcode GTGAAGGTAA Osnat6  
barcode TGTTGTGGTA Osnat7  
barcode GTTGATGAGT Osnat8  
barcode GGTCAGTGTA Osnat9  
barcode GTAATGGAGT Osnat10  
barcode CTCGTTATTC Osnat11  
barcode GGAAGTAAGG Osnat12  
barcode CGGTGTGTGT Osnat13  
barcode CGTCTTCTTA Osnat14

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<div class="post-metadata">

### Author: ![westcott](https://yyz2.discourse-cdn.com/flex036/user_avatar/forum.mothur.org/westcott/32/18_2.png) [@westcott](https://forum.mothur.org/u/westcott)
#### Post date: [November 10, 2014, 7:14pm UTC](https://forum.mothur.org/t/problems-after-trim-seqs/2081/7 "2014-11-10T19:14:03Z")

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Are you using the pdiffs and bdiffs parameters? Can you post a few sequences?
